LOCUS       sequence-419664-        1805 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  In vitro expression of FETCH reporter from the T7 promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 1805)
  AUTHORS   Willi JA, Karim AS, Jewett MC
  TITLE     Cell-Free Translation Quantification via a Fluorescent Minihelix.
  JOURNAL   ACS Synth Biol. 2024 Jul 19;13(7):2253-2259. doi: 
            10.1021/acssynbio.4c00266. Epub 2024 Jul 9.
  PUBMED    38979618
REFERENCE   2  (bases 1 to 1805)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1021/acssynbio.4c00266"; journalName: "ACS Synth Biol"; date: 
            "2024-07-19- 19"; volume: "13"; issue: "7"; pages: "2253-2259"
COMMENT     Sequence Label: pJL1-FETCH2
FEATURES             Location/Qualifiers
     source          1..1805
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      17..605
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     506..525
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     633..652
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        633..651
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             683..705
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             725..742
                     /codon_start=1
                     /product="tetracysteine peptide that binds biarsenical 
                     labeling reagents"
                     /label=tetracysteine tag
                     /translation="CCPGCC"
     primer_bind     complement(806..824)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      820..867
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     CDS             complement(885..1700)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSKPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     1608..1627
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1692..1711
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
ORIGIN
        1 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
       61 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
      121 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
      181 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
      241 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
      301 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
      361 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
      421 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
      481 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
      541 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
      601 ggaaaaacgc cagcaacgcg atcccgcgaa attaatacga ctcactatag ggagaccaca
      661 acggtttccc tctagaaata attttgttta actttaagaa ggagatatac atatgcatcg
      721 ctggtgctgt cctggctgtt gcaagacctt ctaagtcgac cggctgctaa caaagcccga
      781 aaggaagctg agttggctgc tgccaccgct gagcaataac tagcataacc ccttggggcc
      841 tctaaacggg tcttgagggg ttttttgctg aaagccaatt ctgattagaa aaactcatcg
      901 agcatcaaat gaaactgcaa tttattcata tcaggattat caataccata tttttgaaaa
      961 agccgtttct gtaatgaagg agaaaactca ccgaggcagt tccataggat ggcaagatcc
     1021 tggtatcggt ctgcgattcc gactcgtcca acatcaatac aacctattaa tttcccctcg
     1081 tcaaaaataa ggttatcaag tgagaaatca ccatgagtga cgactgaatc cggtgagaat
     1141 ggcaaaagct tatgcatttc tttccagact tgttcaacag gccagccatt acgctcgtca
     1201 tcaaaatcac tcgcatcaac caaaccgtta ttcattcgtg attgcgcctg agcgagacga
     1261 aatacgcgat cgctgttaaa aggacaatta caaacaggaa tcgaatgcaa ccggcgcagg
     1321 aacactgcca gcgcatcaac aatattttca cctgaatcag gatattcttc taatacctgg
     1381 aatgctgttt tcccggggat cgcagtggtg agtaaccatg catcatcagg agtacggata
     1441 aaatgcttga tggtcggaag aggcataaat tccgtcagcc agtttagtct gaccatctca
     1501 tctgtaacat cattggcaac gctacctttg ccatgtttca gaaacaactc tggcgcatcg
     1561 ggcttcccat acaatcgata gattgtcgca cctgattgcc cgacattatc gcgagcccat
     1621 ttatacccat ataaatcagc atccatgttg gaatttaatc gcggcttcga gcaagacgtt
     1681 tcccgttgaa tatggctcat aacacccctt gtattactgt ttatgtaagc agacagtttt
     1741 attgttcatg atgatatatt tttatcttgt gcaatgtaac atcagagatt ttgagacaca
     1801 acgtg
//
